蛇床子素对新生大鼠的心脏保护作用及机制研究  

Cardioprotective effect and mechanism of osthole on neonatal rats

作　　者：肖遥[1] 方丽 周建华[1] 许方雷 杨师午 XIAO Yao;FANG Li;ZHOU Jianhua;XU Fanglei;YANG Shiwu(Department of Cardiovascular Medicine,Hubei Traditional Chinese Medicine Hospital,Wuhan 430061,China;Department of Psychology,Hubei China Resources WISCO Second Staff Hospital,Wuhan 430080,China;Hubei University of Chinese Medicine,Wuhan 430065,China)

机构地区：[1]湖北省中医院心血管内科,湖北武汉430061 [2]湖北华润武钢第二职工医院精神心理科,湖北武汉430080 [3]湖北中医药大学,湖北武汉430065

出　　处：《沈阳药科大学学报》2021年第9期916-923,共8页Journal of Shenyang Pharmaceutical University

基　　金：湖北省科技厅科技条件资源开发项目(2015BCE079)。

摘　　要：目的探究蛇床子素缓解心肌低氧损伤从而对新生大鼠心脏起保护的作用及机制。方法建立新生大鼠常压窒息模型,10、20、40 mg·kg^(-1)蛇床子素治疗。超声心动图检测心脏功能,HE染色观察心肌病理改变,检测肌钙蛋白(cardiac troponin I,cTnI),肌酸激酶同工酶MB(creatine kinase MB,CK-MB),乳酸脱氢酶(lactate dehydrogenase,LDH)和肌红蛋白(myohemoglobin,Mb)浓度,以及丙二醛(malondialdehyde,MDA)和活性氧(reactive oxygen species,ROS)含量,免疫组化检测心肌组织增殖标记蛋白Ki67和半胱天冬酶3(Caspase-3)阳性细胞率,逆转录-聚合酶链式反应(reverse transcription-polymerase chain reaction,RT-PCR)检测鼠双微体基因2(murine double minute 2,MDM2)和p53表达,Western印迹法检测MDM2、p53、蛋白激酶B(protein kinase B,AKT),核因子κB p65(Nuclear factorκB p65),核因子E2相关因子2(nuclear factor E2 related factor2,Nrf2)蛋白表达。结果与正常对照组相比,模型组心肌组织Ki67阳性细胞率降低,Caspase-3阳性细胞率增加,MDM2水平降低,p53水平升高,心肌病理改变加重,心功能减弱,cTnI、CK-MB、Mb、LDH、MDA和ROS水平升高,AKT、NF-κB p65磷酸化比率升高,Nrf2磷酸化比率降低,蛇床子素处理可明显减弱上述各项指标变化,Nrf2抑制剂可扭转蛇床子素心脏保护功能。结论蛇床子素可通过缓解心肌低氧损伤所致心肌细胞病理性凋亡以及氧化应激,对新生大鼠心脏起保护作用。Objective To investigate the effect and mechanism of osthole on relieving myocardial hypoxic injury and protecting the heart of newborn rats.Methods Asphyxia model of neonatal rat was established and treated with 10,20,40 mg·kg^(-1) osthole.Echocardiography was used to detect cardiac function.H&E staining was used to observe myocardial pathological changes, cardiac troponin(cTnI),creatine kinase isoenzymes(CK-MB),lactate dehydrogenase(LDH) and myoglobin(Mb).The melondialdehyde(MDA) and reactive oxygen species(ROS) were measured;The immunohistochemistry was used to detect myocardial tissue proliferation marker protein Ki67 and Caspase-3 positive cell rate;RT-PCR was used to determine the gene expressions of microsomal gene 2(MDM2) and p53;the protein levels of MDM2,p53,protein kinase B(AKT),NF-κB p65 and nuclear factor E2 related factor 2(Nrf2) were detected by Western Blot.Results Compared with control group, the Ki67 positive cell rate of Ki67 in model group was decreased;Caspase-3 positive cell rate was increased;the gene and protein level of MDM2 was decreased;and the gene and protein level of p53 was increased;myocardial pathological changes were aggravated;and cardiac function was decreased.The cTnI,CK-MB,Mb, LDH,MDA and ROS levels were increased, AKT and NF-κB p65 phosphorylation ratios were increased, and Nrf2 phosphorylation ratio was decreased.After osthole treatment, the changes of above indicators were significantly weakened, and Nrf2 inhibitor could reverse the protective effect of osthole.Conclusion Osthol can protect the neonatal rat heart by alleviating the pathological apoptosis and oxidative stress of myocardial cells caused by myocardial hypoxia injury.

关 键 词：新生儿窒息 心肌低氧损伤 蛇床子素 氧化应激 

分 类 号：R285[医药卫生—中药学] R96[医药卫生—中医学]